Characteristics of cerebrospinal fluid oligoclonal band in anti-myelin oligodendrocyte glycoprotein (MOG) antibody associated disease.

Objective: To analyze the immune parameters of cerebrospinal fluid (CSF) and oligoclonal band (OCB) type in patients with anti-myelin oligodendrocyte glycoprotein (MOG) antibody-associated diseases (MOGAD). Methods: Patients who were seropositive for MOG-IgG and diagnosed with MOGAD according to the diagnosis criteria in the Department of Neurology, Huashan Hospital, Fudan University from December 2020 to June 2022 were retrospectively included in this study. Complete clinical data, blood and cerebrospinal fluid samples were collected from all the participants. Paired serum and CSF MOG-IgG and autoimmune encephalitis antibody were assayed by Cell Based Assay (CBA) based on transfected target antigens. Paired serum and CSF albumin and IgG were detected by turbidimetric scattering method, and OCB was detected by standard operation procedure as described. Results: A total of 86 patients (44 males and 42 females) with MOGAD were included in this study, with a median age of 30 years (range: 5-82 years). Among all the patients, 73 patients showed OCB type I, 12 patients showed OCB type II, and one patient showed OCB type III. The overall positive rate of CSF-OCB in MOGAD patients was 15.1 %. The 24-h intrathecal synthesis rate of CSF in the OCB-positive group (n = 13) was higher than that in the OCB-negative group [n = 73, 0.62 (0.26) vs 5.11 (13.67), P = 0.003]. Subgroup analysis revealed that the positive rates of CSF-OCB in the single MOG group (n = 61) and the group combined with other antibodies (n = 25) were 14.8 % and 16.0 %, respectively. The incidence of meningoencephalitis (13/61 vs 13/25, P = 0.011) was significantly different between the two groups. The proportion of patients with high (≥1:32) or low (≤1:10) CSF MOG-IgG also showed significant difference in the group combined with other antibodies (P = 0.032). Optic neuritis was more common in the relapse course group (n = 49) than the monophasic course group (n = 37, P < 0.001) No significant diferences of CSF immune parameters were found in the MOG-IgGserum+/CSF- group and the MOG-IgGserum+/CSF + group, and the titer of MOG-IgG in the serum or CSF did not influence CSF immune parameters in different subgroups. Conclusion: The overall positive rate of CSF-OCB in MOGAD patients was 15.1 %. The 24-h intrathecal synthesis rate of cerebrospinal fluid in the OCB-positive group was higher than that in the OCB-negative group. This study illustrated OCB characterization in MOGAD patients, and will shed light on the standardization of OCB test in the study of immune diseases.

A novel homozygous mutation in TREM2 found in a Chinese early-onset dementia family with mild bone involvement.

Variants in triggering receptor expressed on myeloid cells 2 (TREM2) are associated with both behavioral variant frontotemporal lobar degeneration and Alzheimer's disease. TREM2 homozygous mutations cause Nasu-Hakola disease, an early-onset recessive form of dementia preceded by bone cysts and fractures. The same type of mutations has been shown to cause frontotemporal dementia without the presence of any bone phenotype. Herein, we report a Chinese Han consanguineous family carrying a novel TREM2 mutation, presenting with early-onset dementia similar to behavioral variant frontotemporal dementia with mild radiological bone involvement. Minigene reporter assay showed the variant disturbed splicing by preservation of intron 2 in transcription. In our investigation, the clinical and genetic spectra of Chinese early-onset dementia patients were expanded; TREM2 mutations should be screened in familial and Chinese early-onset dementia patients.

HLA typing using next-generation sequencing for Chinese juvenile- and adult-onset myasthenia gravis patients.

To compare HLA typing between juvenile- and adult-onset myasthenia gravis (MG), we enrolled 101 children (age ≤12 years) and 168 adults (age ≥20 years) with MG. We excluded patients with histories of thymoma, thyroid disease, or other autoimmune disease. We selected 41 seronegative juvenile-onset patients with ocular symptoms only, and 41 seropositive adult-onset patients with generalized symptoms. We used next-generation sequencing for typing and analysis of HLA genes (Loci: A, B, C, DPA1, DPB1, DQA1, DQB1 and DRB1). Haplotypes HLA-A∗02:07:01-B∗46:01:01-C∗01:02:01-DQA1∗01:01:01-DQB1∗03:03:02-DRB1∗09:01:02, HLA-A∗11:01:01, HLA-A∗24:02:01, and HLA-DPA1∗02:02:02 were found to be related to juvenile-onset MG and HLA-A∗01:01:01, HLA-A∗02:03:01, HLA-C∗03:04:01, and HLA-DQB1∗06:02:01 to adult-onset MG. Therefore, our findings suggested that HLA typing might determine the heterogeneity between AChR-Ab negative juvenile-onset and AChR-Ab positive adult-onset Chinese MG patients.

ß-Amyloid Upregulates Intracellular Clusterin but not Secretory Clusterin in Primary Cultured Neurons and APP Mice.

BACKGROUND: Previous studies have suggested that the expression of Aß and clusterin is positively correlated. However, the causal relationship between Aß and clusterin has not been exactly clarified. METHODS: In this study, primary hippocampal neurons were treated with Aß42; clusterin mRNA and protein expression was assessed. Furthermore, we evaluated Aß and clusterin protein expression in the brains of APP/PSEN1 mice, as well as serum clusterin concentration. RESULTS: We observed here that the exposure of primary hippocampal neurons to Aß42 induced an overexpression of intracellular clusterin, but the level of clusterin in supernatants was not changed. Moreover, in APP/PSEN1 mice, there was a significant increase in intracellular clusterin in cortex and hippocampus, compared to age-matched WT mice, while serum clusterin level in APP/PSEN1 mice and in WT mice has no significant difference. CONCLUSION: Aß42 upregulated intracellular clusterin, but secretory clusterin did not change. These findings reveal that clusterin is upregulated by Aß and is responsive to AD pathology, although plasma clusterin concentration is not evidenced to be a stand-alone biomarker for AD.

[Levels of histone H3 acetylation in peripheral blood mononuclear cells of acute cerebral infarction patients].

OBJECTIVE: To observe the co-variation of acetylated histone H3 levels of peripheral blood mononuclear cells (PBMCs) and onset time of acute ischemic stroke and examine the histone H3 acetylation levels in PBMCs of acute cerebral infarction patients with different stroke subtypes and injury degrees. METHODS: The peripheral blood samples 2 ml from patients at different timepoints (1, 3, 5, 7, 14 d) from April 2013 to July 2013 and normal controls were collected to observe the dynamic change of Ac-H3 levels of PBMCs in cerebral infarction patients. Also between April 2013 and October 2013, blood samples from 103 patients within 7 d after acute ischemic stroke were collected. Global histone was extracted by assay kit and differential histone H3 acetylation levels were determined by Western blot. All patients were measured by the Oxfordshire Community Stroke Project (OCSP) classification and National Institutes of Health Stroke Scale (NIHSS) score. RESULTS: The levels of acetylated histone H3 in PBMCs of acute cerebral infarction patients started to decrease as early as 1 d and remained below those normal controls for at least 7 d after stroke. It fulfilled the minimum at 3 d after infarction (P < 0.001). Acetylated histone H3 levels of PBMCs differed in OCSP classification (P < 0.05) and reached a nadir in TACI group. Histone H3 acetylation levels of PBMCs were major affecting factors of neurological injury severity and negatively correlated with it through multiple regressive analysis (ß = -0.297, P = 0.001). CONCLUSION: Histone H3 acetylation level in PBMCs of acute cerebral infarction patients is lower than healthy persons. And it decreases markedly in TACI group and patients with severe neurological dysfunction.

[Acupuncture treatment programs for post-stroke motor rehabilitation in community hospitals: study protocol of a multicenter, randomized, controlled trial].

BACKGROUND: Stroke is responsible for increasingly high rates of mortality and disability worldwide. Approximately two million people suffer from stroke for the first time in China each year. The high incidence (50%) of post-stroke disability brings a heavy burden to patients and their caregivers. Acupuncture has been widely used in the communities for post-stroke rehabilitation in China. The objective of this trial is to apply our acupuncture research achievement to treatment and evaluation of post-stroke hemiplegic patients in community. METHODS AND DESIGN: A multicenter, randomized, controlled trial will be performed in Longhua Hospital and a number of community health service centers in Shanghai. A total of 124 patients (estimated sample size) with post-stroke hemiplegia will be randomly divided into an acupuncture group and a control group. The patients undergoing randomization should be stratified according to National Institutes of Health Stroke Scale score at baseline. Within the acupuncture group, different acupuncture protocols are administered to patients with flaccid paralysis or spastic paralysis based on the Ashworth Scale. Patients in the acupuncture group will also be treated with comprehensive rehabilitation therapy. The control group will be treated with comprehensive rehabilitation therapy only. The primary outcome measures are the Simplified Fugl-Meyer Motor Scale, the Modified Barthel Index, and the Burden of Stroke Scale. Secondary outcome measures are the modified Rankin Scale, the modified Ashworth Scale and the Stroke Scale of Traditional Chinese Medicine. Outcome measures will be performed after 4 and 8 weeks of treatment. The patients will be followed up after 6 months. DISCUSSION: The results of this study are expected to demonstrate that our standardized acupuncture protocol for treating and evaluating post-stroke hemiplegic patients will improve motor function and lessen the burden of post-stroke patients within the communities. This will provide the evidence to support successful translation of acupuncture therapy for post-stroke hemiplegic patients in community hospital use. TRIAL REGISTRATION: This trial was registered in Chinese Clinical Trial Registry with the registration number ChiCTR-TRC-11001347.

Pretreatment with PTD-calbindin D 28k alleviates rat brain injury induced by ischemia and reperfusion.

Calcium toxicity remains the central focus of ischemic brain injury. Calcium channel antagonists have been reported to be neuroprotective in ischemic animal models but have failed in clinical trials. Rather than block the calcium channels, calbindin proteins can buffer excessive intracellular Ca2+, and as a result, maintain the calcium homeostasis. In the present study, we investigated the effect of calbindin D 28k (CaBD) in ischemic brain using the novel technique protein transduction domain (PTD)-mediated protein transduction. We generated PTD-CaBD in Escherichia coli, tested its biologic activity in N-methyl-D-aspartate (NMDA)- and oxygen-glucose deprivation (OGD)-induced hippocampal injury models, and examined the protection of the fusion protein using a rat brain focal ischemia model. Infarct volume was determined using 2,3,5-triphenyl-tetrazolium chloride staining; neuronal injury was examined using terminal deoxynucleotidyl transferase-mediated 2'-deoxyuridine 5'-triphosphate-biotin nick end labeling (TUNEL) staining and cleaved caspase-3 assay. The results showed that the PTD-CaBD was efficiently delivered into Cos7 cells, hippocampal slice cells, and brain tissue. Pretreatment with PTD-CaBD decreased intracellular free calcium concentration and reduced cell death in NMDA- or OGD-exposed hippocampal slices (P<0.05). Intraperitoneal administration of PTD-CaBD before transient middle cerebral artery occlusion decreased brain infarct volume (280+/-47 versus 166+/-70 mm3, P<0.05), and improved neurologic outcomes compared with the control. Further studies showed that, compared with the control animals, PTD-CaBD decreased TUNEL (58%+/-7% versus 29%+/-3%, P<0.05)- and cleaved caspase-3 (62+/-4/field versus 31+/-6/field, P<0.05)-positive cells in the ischemic boundary zone. These results indicate that systemic administration of PTD-CaBD could attenuate ischemic brain injury, suggesting that PTD-mediated protein transduction might provide a promising and effective approach for the therapies of brain diseases, including cerebral ischemia.

Quantification of huperzine A in Huperzia serrata by HPLC-UV and identification of the major constituents in its alkaloid extracts by HPLC-DAD-MS-MS.

A rapid, simple and reliable high performance liquid chromatography (HPLC) method has been established for the analysis of the major alkaloids in Huperzia serrata, a traditional Chinese medicine (TCM) herb. After chromatographic separation on a reversed-phase C18 column with methanol-ammonium acetate (pH 6.0; 80 mM, 30/70, v/v) as the mobile phase, nine alkaloid compounds in the alkaloid extracts of H. serrata were identified by online diode array detection-MS and by comparing with data from literature and standard samples. One compound in the extract, huperzine A, is a drug for treating Alzheimer's disease. Its content was quantified by HPLC coupled with UV-vis. The method was the validated. The recovery rates were 96.8-97.7% with R.S.D <2.44%. The intra- and inter-day precisions, expressed as R.S.D., ranged from 0.53% to 1.51%. Good linear regression was observed in the concentration range of 5-100 microg/ml (r = 0.9997). The results demonstrate that this method is simple, selective, and suitable for the quality control of this TCM herb.